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In cancer, human cells lose the ability to properly control the series of events that occur constantly during cell growth and division, including protein expression, stability, and dynamics. Heat shock proteins (Hsps) are key molecules in these events, constitutively expressed at high levels and could furthermore be induced by the response to cancer-induced stress. In tumor cells, Hsps have been shown to be implicated in the regulation of apoptosis, immune responses, angiogenesis and metastasis; in some cases, they can be overexpressed and dysregulated, representing important cancer hallmarks. In the past few years, it has been demonstrated that Hsps can be released by tumor cells through several secreting pathways, including the extracellular vesicles (EVs), thus modulating the tumor microenvironment as well as long-distance intercellular communication and metastatization. In this review, we discuss the role of extracellular Hsps in cancer, with a particular interest in Hsps in EVs. We would also like to highlight the importance of fully understanding of the role of extracellular Hsps released by EVs and encourage further research in this field the use of Hsps as early cancer biomarkers and therapeutic targets.

In human studies, frustration has been identified as a potential risk factor for substance use disorders. Currently, there is little research into the role of frustration in substance use disorders despite research showing that frustration tolerance in humans is associated with a lower likelihood of developing substance use problems, better outcomes in recovery, and fewer relapses. To address this need, our studies use rat self-administration models to focus on frustration-related behavior in natural reward and addiction-related behavioral procedures. First, to study frustration in operant responding, there is a need to establish a real-time objective measure and validate its use in predicting vulnerability to drug use. Frustration is when a subject cannot achieve a reinforcer, receives less than the anticipated reinforcer, or has to work harder to achieve a reinforcer. Therefore, the measure of frustration should increase in instances consistent with the definition of frustration. Furthermore, the operant measure of frustration should assess a form of either the approach or avoidance responses to frustration. Increases in bar press durations are shown to be an approach strategy that can be used to measure frustration-related behavior objectively and, in some conditions, has the potential to signal a future transition to avoidance strategies. Supplementary experiments solidify that changes in bar press durations are a modification of continued approach behavior that can be used to objectively measure frustration-related behavior by satisfying nine behavioral criteria for barpress durations to measure frustration. Additional experiments established the predictive validity of identifying individual differences in frustration-like behavior as measured by bar press durations prior to drug self-administration. This work also affirms bar press durations as the optimal measure of frustration-like behavior within operant self-administration compared to the force of a bar press as a potential alternative approach response variation. Essentially, the results within this dissertation solidify bar press durations as an effective measure of frustration-like behavior to further elucidate the understanding of frustration within substance use disorders.

The purpose of this preclinical study was to evaluate the influence of tamoxifen (TAM) on the peri-implant bone remodeling of osseointegrated titanium implants in ovariectomized female rats.Seventy-two female rats underwent bilateral ovariectomy 20 weeks before implants placement. One titanium implant was inserted in each tibia of the animals. Six weeks following the implant surgery, animals were randomly divided into two experimental groups (n = 36), which received either saline solution (SS) or tamoxifen citrate (TAM) via gavage until euthanasia. Euthanasia was performed at 30, 60, and 90 days after the first gavage. Assessments of bone to implant contact (BIC), bone ingrowth percentage (BIN), morphological description of cellular and tissue reactions, immunohistochemistry for the detection of bone morphogenetic protein 2/4 (BMP2/4), runt-related transcription factor 2 (RUNX-2), osteocalcin (OCN) and tartrate-resistant acid phosphatase (TRAP), and bone chemical composition through scanning electron microscopy with energy-dispersive x-ray spectroscopy were performed.Tamoxifen group presented higher BIC, higher BIN, higher RUNX-2 and OCN, lower TRAP-positive cells/mmTamoxifen enhanced the remodeling of the bone surrounding titanium implants in ovariectomized rats.

Reference data on the asymmetry of facial thirds of children is still scarce, although it can offer meaningful comparative information for clinical studies. This study aims to provide reference data on the facial asymmetry of Italian children using a 3D analysis of facial thirds divided according to the trigeminal nerve distribution (upper, middle, and lower). A 3D surface-based approach was conducted on the digital models of the faces of 135 children (74 M, 61 F), acquired by stereophotogrammetry. In addition to sex, two different age classes were analyzed (4–8 years and 9–12 years). For each facial third, the asymmetry was expressed as root-mean-square distance (RMS) by calculating the point-to-point distances between the original and the reflected 3D models. A 3-way ANOVA test verified significant differences between the two sexes, the two age classes, and the three facial thirds (p-value < 0.05), and also their interaction. Significant differences were found between the sexes (females were more symmetric, p = 0.005) and the two age groups (younger were more symmetric p < 0.001). According to Tukey’s HSD post-hoc test, among the thirds, the middle one proved to be significantly more symmetrical (p < 0.001). No significant interaction impacting the asymmetry was found when the main factors were considered in any combination. Reference data on the “normal” facial asymmetry of Italian children was provided for further clinical purposes. Normal children have low average RMS values (0.30–0.51 mm) and younger, in particular females, proved more symmetrical than older children, while among the facial surfaces, the middle proved most symmetrical in both sexes, although with little clinical relevance. Since there is no consensus on the amount of symmetry deemed clinically acceptable, further studies on larger randomized samples are auspicial.

The currently available nebulization devices have a slow aerosol flow and produce vapor with large microdrops. Improved devices that achieve higher airflow and produce smaller microdrops are needed to improve the clinical care of patients. To address this critical need, we developed a novel system for the molecular vaporization of liquids. This device vaporizes an active pharma-cological substance dissolved in water, alcohol, or a mixture of water and alcohol using two energy sources at the same time: high-frequency ultrasound and thermal induction. Application of energy to a solution contained in the device's tank allows, within tens of seconds, for the vaporization of the solution itself, with the generation of a vapor consisting of microdrops of very small diameter (0.2-0.3 mu m). In this article, we illustrate the technology used, the main verifi-cation tests performed, and the primary fields of application for this device. In particular, the advantages of both the aerosol delivery system and the administration system are highlighted.

(1) Background: Atopic dermatitis is one of the most common inflammatory skin diseases characterized by T helper (Th) 2 and Th22 cells producing interleukin (IL)-4/IL-13 and IL-22, respectively. The specific contribution of each cytokine to the impairment of the physical and the immune barrier via Toll-like receptors (TLRs) is poorly addressed concerning the epidermal compartment of the skin. (2) Methods: The effect of IL-4, IL-13, IL-22, and the master cytokine IL-23 is evaluated in a 3D model of normal human skin biopsies (n = 7) at the air–liquid interface for 24 and 48 h. We investigated by immunofluorescence the expressions of (i) claudin-1, zonula occludens (ZO)-1 filaggrin, involucrin for the physical barrier and (ii) TLR2, 4, 7, 9, human beta-defensin 2 (hBD-2) for the immune barrier. (3) Results: Th2 cytokines induce spongiosis and fail in impairing tight junction composition, while IL-22 reduces and IL-23 induces claudin-1 expression. IL-4 and IL-13 affect the TLR-mediated barrier largely than IL-22 and IL-23. IL-4 early inhibits hBD-2 expression, while IL-22 and IL-23 induce its distribution. (4) Conclusions: This experimental approach looks to the pathogenesis of AD through molecular epidermal proteins rather than cytokines only and paves the way for tailored patient therapy.

Mutations in genes encoding molecular chaperones, for instance the genes encoding the subunits of the chaperonin CCT (chaperonin containing TCP-1, also known as TRiC), are associated with rare neurodegenerative disorders. Using a classical molecular dynamics approach, we investigated the occurrence of conformational changes and differences in physicochemical properties of the CCT5 mutations His147Arg and Leu224Val associated with a sensory and a motor distal neuropathy, respectively. The apical domain of both variants was substantially but differently affected by the mutations, although these were in other domains. The distribution of hydrogen bonds and electrostatic potentials on the surface of the mutant subunits differed from the wild-type molecule. Structural and dynamic analyses, together with our previous experimental data, suggest that genetic mutations may cause different changes in the protein-binding capacity of CCT5 variants, presumably within both hetero- and/or homo-oligomeric complexes. Further investigations are necessary to elucidate the molecular pathogenic pathways of the two variants that produce the two distinct phenotypes. The data and clinical observations by us and others indicate that CCT chaperonopathies are more frequent than currently believed and should be investigated in patients with neuropathies.

We present a thorough structural characterization of Graphene Nano Particles (GNPs) prepared by means of physical procedures, i.e., ball milling and ultra-sonication of high-purity synthetic graphite. UV-vis absorption/extinction spectroscopy, Dynamic Light Scattering, Transmission Electron Microscopy, IR and Raman spectroscopies were performed. Particles with small size were obtained, with an average lateral size = 70–120 nm, formed by few = 1–10 stacked layers, and with a small number of carboxylic groups on the edges. GNPs relatively more functionalized were separated by centrifugation, which formed stable water dispersions without the need for any surfactant. A critical reading and unified interpretation of a wide set of spectroscopic data was provided, which demonstrated the potential of Specular Reflectance Infrared Spectroscopy for the diagnosis and quantification of chemical functionalization of GNPs. Raman parameters commonly adopted for the characterization of graphitic materials do not always follow a monotonic trend, e.g., with the particle size and shape, thus unveiling some limitations of the available spectroscopic metrics. This issue was overcome thanks to a comparative spectra analysis, including spectra deconvolution by means of curve fitting procedures, experiments on reference materials and the exploitation of complementary characterization techniques.

Liver resection is the best treatment for hepatocellular carcinoma (HCC) when resectable. Unfortunately, many patients with HCC cannot undergo liver resection. Percutaneous thermoablation represents a valid alternative for inoperable neoplasms and for small HCCs, but it is not always possible to accomplish it. In cases where the percutaneous approach is not feasible (not a visible lesion or in hazardous locations), laparoscopic thermoablation may be indicated. HCC diagnosis is commonly obtained from imaging modalities, such as CT and MRI, However, the interpretation of radiological images, which have a two-dimensional appearance, during the surgical procedure and in particular during laparoscopy, can be very difficult in many cases for the surgeon who has to treat the tumor in a three-dimensional environment. In recent years, more technologies have helped surgeons to improve the results after ablative treatments. The three-dimensional reconstruction of the radiological images has allowed the surgeon to assess the exact position of the tumor both before the surgery (virtual reality) and during the surgery with immersive techniques (augmented reality). Furthermore, indocyanine green (ICG) fluorescence imaging seems to be a valid tool to enhance the precision of laparoscopic thermoablation. Finally, the association with laparoscopic ultrasound with contrast media could improve the localization and characteristics of tumor lesions. This article describes the use of hepatic three-dimensional modeling, ICG fluorescence imaging and laparoscopic ultrasound examination, convenient for improving the preoperative surgical preparation for personalized laparoscopic approach.

Despite recent advances in understanding the causes of epilepsy, especially the genetic, comprehending the biological mechanisms that lead to the epileptic phenotype remains difficult. A paradigmatic case is constituted by the epilepsies caused by altered neuronal nicotinic acetylcholine receptors (nAChRs), which exert complex physiological functions in mature as well as developing brain. The ascending cholinergic projections exert potent control of forebrain excitability, and wide evidence implicates nAChR dysregulation as both cause and effect of epileptiform activity. First, tonic-clonic seizures are triggered by administration of high doses of nicotinic agonists, whereas non-convulsive doses have kindling effects. Second, sleep-related epilepsy can be caused by mutations on genes encoding nAChR subunits widely expressed in the forebrain (CHRNA4, CHRNB2, CHRNA2). Third, in animal models of acquired epilepsy, complex time-dependent alterations in cholinergic innervation are observed following repeated seizures. Heteromeric nAChRs are central players in epileptogenesis. Evidence is wide for autosomal dominant sleep-related hypermotor epilepsy (ADSHE). Studies of ADSHE-linked nAChR subunits in expression systems suggest that the epileptogenic process is promoted by overactive receptors. Investigation in animal models of ADSHE indicates that expression of mutant nAChRs can lead to lifelong hyperexcitability by altering i) the function of GABAergic populations in the mature neocortex and thalamus, ii) synaptic architecture during synaptogenesis. Understanding the balance of the epileptogenic effects in adult and developing networks is essential to plan rational therapy at different ages. Combining this knowledge with a deeper understanding of the functional and pharmacological properties of individual mutations will advance precision and personalized medicine in nAChR-dependent epilepsy.

Background Triple-negative breast cancer (TNBC) is the most heterogeneous and malignant subtype of breast cancer (BC). TNBC is defined by the absence of expression of estrogen, progesterone and HER2 receptors and lacks efficacious targeted therapies. NEK2 is an oncogenic kinase that is significantly upregulated in TNBC, thereby representing a promising therapeutic target. NEK2 localizes in the nucleus and promotes oncogenic splice variants in different cancer cells. Notably, alternative splicing (AS) dysregulation has recently emerged as a featuring trait of TNBC that contributes to its aggressive phenotype. Methods To investigate whether NEK2 modulates TNBC transcriptome we performed RNA-sequencing analyses in a representative TNBC cell line (MDA-MB-231) and results were validated in multiple TNBC cell lines. Bioinformatics and functional analyses were carried out to elucidate the mechanism of splicing regulation by NEK2. Data from The Cancer Genome Atlas were mined to evaluate the potential of NEK2-sensitive exons as markers to identify the TNBC subtype and to assess their prognostic value. Results Transcriptome analysis revealed a widespread impact of NEK2 on the transcriptome of TNBC cells, with 1830 AS events that are susceptible to its expression. NEK2 regulates the inclusion of cassette exons in splice variants that discriminate TNBC from other BC and that correlate with poor prognosis, suggesting that this kinase contributes to the TNBC-specific splicing program. NEK2 elicits its effects by modulating the expression of the splicing factor RBFOX2, a well-known regulator of epithelial to mesenchymal transition (EMT). Accordingly, NEK2 splicing-regulated genes are enriched in functional terms related to cell adhesion and contractile cytoskeleton and NEK2 depletion in mesenchymal TNBC cells induces phenotypic and molecular traits typical of epithelial cells. Remarkably, depletion of select NEK2-sensitive splice-variants that are prognostic in TNBC patients is sufficient to interfere with TNBC cell morphology and motility, suggesting that NEK2 orchestrates a pro-mesenchymal splicing program that modulates migratory and invasive properties of TNBC cells. Conclusions Our study uncovers an extensive splicing program modulated by NEK2 involving splice variants that confer an invasive phenotype to TNBCs and that might represent, together with NEK2 itself, valuable therapeutic targets for this disease.

This essay's main goal the present review is to highlight the connections between intestinal dysbiosis and the ensuing activation of the mucosal lymphatic system. One of the study's goals is to investigate the impact on mood caused by a serotonergic deficit driven by mucosal inflammation. It assesses the relationship between food consumption and the onset of psychological and mental illness as a secondary end aim. Patients with inflammatory bowel diseases and psychological and psychiatric mood disorders appear to benefit therapeutically from the sort of diet they consume.

Burn wound healing is a very intricate and complex process that conventionally includes three interrelated and overlapping stages of hemostasis/inflammation, proliferation and remodeling. This review aims to explore the molecular interactions of NGF with the most prominent cell types in the skin and their respective secretory products during wound healing, particularly burn wound healing. Different types of cells such as, nerve cells, endothelial cells, mast cells, macrophages, neutrophils, keratinocytes and fibroblasts all come into play through a plethora of cytokines and growth factors including nerve growth factor (NGF). NGF is a pleiotropic molecule that exerts its effects on all the aforementioned cells using two types of receptors (TrkA and p75) and affects wound healing by decreasing healing time and improving the quality of the scar. Both receptors mediate cellular proliferation, survival and apoptosis through complex signaling molecules. During the inflammatory phase, macrophages and mast cells secrete ample cytokines and growth factors, including NGF, which participate in the inflammatory reaction and induction of other cells targeting a homeostatic state. The proliferative phase follows, and NGF promotes angiogenesis through VEGF and FGF expression in endothelial cells. NGF also stimulates keratinocyte proliferation and neurite extension through the TrkA-PI3K/Akt pathway. Other molecules such as TGF-β1, IL-1β and TNF-α increase NGF expression in fibroblasts through dynamic interactions with Smads and MAPK molecules. Stimulated fibroblasts induce new collagen production to form the granulation tissue. In the remodeling phase, NGF regulates fibroblasts and induces their differentiation into myofibroblasts ultimately leading to wound contracture. In addition, NGF stimulates melanocytes and enhances hair growth and pigmentation. Such data depict the mechanisms of action of NGF implicated in the various stages of the healing process and support its applicability as a new targeted therapeutic molecule effective in burn wound healing but with some limitations.

Epidermal junctions help to preserve cutaneous homeostasis and, consequently, protect the body against a wide range of environmental stresses [...]

Prostate cancer (PC) relies on androgen receptor (AR) signaling. While hormonal therapy (HT) is efficacious, most patients evolve to an incurable castration-resistant stage (CRPC). To date, most proposed mechanisms of acquired resistance to HT have focused on AR transcriptional activity. Herein, we uncover a new role for the AR in alternative cleavage and polyadenylation (APA). Inhibition of the AR by Enzalutamide globally regulates APA in PC cells, with specific enrichment in genes related to transcription and DNA topology, suggesting their involvement in transcriptome reprogramming. AR inhibition selects promoter-distal polyadenylation sites (pAs) enriched in cis-elements recognized by the cleavage and polyadenylation specificity factor (CPSF) complex. Conversely, promoter-proximal intronic pAs relying on the cleavage stimulation factor (CSTF) complex are repressed. Mechanistically, Enzalutamide induces rearrangement of APA subcomplexes and impairs the interaction between CPSF and CSTF. AR inhibition also induces co-transcriptional CPSF recruitment to gene promoters, predisposing the selection of pAs depending on this complex. Importantly, the scaffold CPSF160 protein is up-regulated in CRPC cells and its depletion represses HT-induced APA patterns. These findings uncover an unexpected role for the AR in APA regulation and suggest that APA-mediated transcriptome reprogramming represents an adaptive response of PC cells to HT.

Inflammatory bowel disease (IBD), including Crohn's disease and ulcerative colitis, are chronic and recurrent inflammatory diseases. Although the typical symptoms are localized in the gastrointestinal tract, there are also extraintestinal manifestations (EIMs) associated with IBD, probably caused by immune reactions secondary to the pathology of origin. Anybody can be affected by EIMs, which can be dermatological, pulmonary, ocular, musculoskeletal, hepatobiliary and neurological, et al. This represents a therapeutic challenge for clinicians, who must first establish a diagnostic and then a multidisciplinary therapeutic path, in order to guarantee an optimal quality of life for the patient.

The aim of the present study was to evaluate if, after treatment with aligners (ALIGN) and fixed orthodontic appliance (FOA), alterations of the neuromuscular activity may occur and if differences in these changes can be detected between the two treatments. Sixteen healthy patients (7 FOA, 9 ALIGN) with class I or class II molar relation were recruited. Standardized surface electromyography (ssEMG) was used to evaluate the activity of the masticatory muscles (masseters-MM and temporalis-TM) before the beginning of the orthodontic treatment (T1), at the end (T2), and 3 months (T3) after the end of the treatment. Intragroup (within timepoints) and intergroup differences were statistically analyzed. At T1, the mean values of each ssEMG index were within the normal range in both groups. At T2, the FOA group showed larger differential recruitment of the MM than TA muscles with a value slightly over the normal range. All the indexes were normalized at T3, and no differences emerged between groups. In the FOA group, the index of MM symmetrical contraction increased significantly at T3 compared to T1 and T2. In the ALIGN group, no significant changes were observed between each timepoint. In FOA subjects, a slight alteration of the muscular activity appeared immediately after bracket removal and this alteration normalized after 3 months of rescue. In subjects treated with aligners, no significant alteration of the muscular activity was assessed.

Inflammatory Bowel diseases (IBDs) constitute a complex panel of disorders characterized with chronic inflammation affecting the alimentary canal along with extra intestinal manifestations. Its exact etiology is still unknown; however, it seems to be the result of uncharacterized environmental insults in the intestine and their immunological consequences along with dysbiosis, in genetically predisposed individuals. It was the main target of our team since 2002 to explore the etiology of IBD and the related role of bacteria. For almost two decades, our laboratory, among others, has been involved in the reciprocal interaction between the host gastrointestinal lining and the homing microbiota. In the first decade, the attention of scientists focused on the possible role of enteropathogenic E. coli and its relationship to the mechanistic pathways involved in IBD induced in both rats and mice by chemicals like Iodoacetamide, Dextran Sodium Sulfate, Trinitrobenzene, thus linking microbial alteration to IBD pathology. A thorough characterization of the various models was the focus of research in addition to exploring how to establish an active homeostatic composition of the commensal microbiota, including its wide diversity by restoration of gut microbiota by probiotics and moving from dysbiosis to eubiosis. In the last six years and in order to effectively translate such findings into clinical practice, it was critical to explore their relationship to colorectal cancer CRC both in solid tumors and chemically induced CRC. It was also critical to explore the degree of intestinal dysbiosis and linking to IBD, CRC and diabetes. Remarkably, the active mechanistic pathways were proposed as well as the role of microbiota or bacterial metabolites involved. This review covers two decades of investigations in our laboratory and sheds light on the different aspects of the relationship between microbiota and IBD with an emphasis on dysbiosis, probiotics and the multiple mechanistic pathways involved.

Alternative splicing and polyadenylation represent two major steps in pre‐mRNA‐processing, which ensure proper gene expression and diversification of human transcriptomes. Deregulation of these processes contributes to oncogenic programmes involved in the onset, progression and evolution of human cancers, which often result in the acquisition of resistance to existing therapies. On the other hand, cancer cells frequently increase their transcriptional rate and develop a transcriptional addiction, which imposes a high stress on the pre‐mRNA‐processing machinery and establishes a therapeutically exploitable vulnerability. A prominent role in fine‐tuning pre‐mRNA‐processing mechanisms is played by three main families of protein kinases: serine arginine protein kinase (SRPK), CDC‐like kinase (CLK) and cyclin‐dependent kinase (CDK). These kinases phosphorylate the RNA polymerase, splicing factors and regulatory proteins involved in cleavage and polyadenylation of the nascent transcripts. The activity of SRPKs, CLKs and CDKs can be altered in cancer cells, and their inhibition was shown to exert anticancer effects. In this review, we describe key findings that have been reported on these topics and discuss challenges and opportunities of developing therapeutic approaches targeting splicing factor kinases., Phosphorylation of RNA polymerase II and regulatory RNA‐binding proteins represents a major modality of pre‐mRNA‐processing regulation. Serine arginine protein kinase (SRPK), CDC‐like kinase (CLK) and cyclin‐dependent kinase (CDK) are the three main families of protein kinases regulating this process. This review reports findings that have highlighted oncogenic alteration of the pre‐mRNA‐processing as a consequence of aberrant regulation of SRPK, CLK and CDK activity and discuss challenges and opportunities of developing therapeutic approaches targeting these kinases.